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Journal of Biomedical Optics

Extended cavity laser enhanced two-photon flow cytometry
Author(s): Eric R. Tkaczyk; Alan H. Tkaczyk; Steve M. Katnik; Jing Yong Ye; Kathryn E. Luker; Gary D. Luker; Andrzej Myc; James R. Baker, Jr.; Theodore B. Norris
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Paper Abstract

We demonstrate enhanced sensitivity in two-photon flow cytometry with an extended cavity laser excitation source. At low power, the home-built 20-MHz oscillator was able to detect a significantly larger fraction, in either phosphate buffered saline (PBS) or whole blood, of green fluorescent protein (GFP)–expressing MCA-207 cells cross-labeled with the membrane-binding lipophilic dye DiD. A geometrical model is used to explain unique features of the signals resulting from the different spatial distribution of DiD and GFP. These unique features include sub-square law scaling of unsaturated two-photon signal, a sigmoidal sensitivity curve for detection under varying powers for cell detection thresholds as low as a single photon, and uncorrelated signal strengths in two detection channels.

Paper Details

Date Published: 1 July 2008
PDF: 12 pages
J. Biomed. Opt. 13(4) 041319 doi: 10.1117/1.2967983
Published in: Journal of Biomedical Optics Volume 13, Issue 4
Show Author Affiliations
Eric R. Tkaczyk, Univ. of Michigan (United States)
Alan H. Tkaczyk, Univ. of Michigan (United States)
Steve M. Katnik, Univ. of Michigan (United States)
Jing Yong Ye, Univ. of Michigan (United States)
Kathryn E. Luker, Univ. of Michigan Medical School (United States)
Gary D. Luker, Univ. of Michigan Medical School (United States)
Andrzej Myc, Univ. of Michigan (United States)
James R. Baker, Jr., Univ. of Michigan (United States)
Theodore B. Norris, Univ. of Michigan (United States)

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